NAD (P) transhydrogenase has vital non‐mitochondrial functions in malaria parasite transmission

Nicotinamide adenine dinucleotide (NAD) and its phosphorylated form (NADP) are vital for cell function in all organisms and form cofactors to a host of enzymes in catabolic and anabolic processes. NAD(P) transhydrogenases (NTHs) catalyse hydride ion transfer between NAD(H) and NADP(H). Membrane-bound NTH isoforms reside in the cytoplasmic membrane of bacteria, and the inner membrane of mitochondria in metazoans, where they generate NADPH. Here, we show that malaria parasites encode a single membrane-bound NTH that localises to the crystalloid, an organelle required for sporozoite transmission from mosquitos to vertebrates. We demonstrate that NTH has an essential structural role in crystalloid biogenesis, whilst its enzymatic activity is required for sporozoite development. This pinpoints an essential function in sporogony to the activity of a single crystalloid protein. Its additional presence in the apicoplast of sporozoites identifies NTH as a likely supplier of NADPH for this organelle during liver infection. Our findings reveal that Plasmodium species have co-opted NTH to a variety of non-mitochondrial organelles to provide a critical source of NADPH reducing power

A Novel Family of Toxoplasma IMC Proteins Displays a Hierarchical Organization and Functions in Coordinating Parasite Division

Apicomplexans employ a peripheral membrane system called the inner membrane complex (IMC) for critical processes such as host cell invasion and daughter cell formation. We have identified a family of proteins that define novel sub-compartments of the Toxoplasma gondii IMC. These IMC Sub-compartment Proteins, ISP1, 2 and 3, are conserved throughout the Apicomplexa, but do not appear to be present outside the phylum. ISP1 localizes to the apical cap portion of the IMC, while ISP2 localizes to a central IMC region and ISP3 localizes to a central plus basal region of the complex. Targeting of all three ISPs is dependent upon N-terminal residues predicted for coordinated myristoylation and palmitoylation. Surprisingly, we show that disruption of ISP1 results in a dramatic relocalization of ISP2 and ISP3 to the apical cap. Although the N-terminal region of ISP1 is necessary and sufficient for apical cap targeting, exclusion of other family members requires the remaining C-terminal region of the protein. This gate-keeping function of ISP1 reveals an unprecedented mechanism of interactive and hierarchical targeting of proteins to establish these unique sub-compartments in the Toxoplasma IMC. Finally, we show that loss of ISP2 results in severe defects in daughter cell formation during endodyogeny, indicating a role for the ISP proteins in coordinating this unique process of Toxoplasma replication

Clostridial infection after open fractures of the lower extremity - report of two cases and discussion of pathomechanism and treatment

Management of post-traumatic open fractures resulting from severe injuries of the lower extremity continues to challenge orthopedic and reconstructive surgeons. Moreover, post-traumatic osteoarticular infections due to Clostridium species are rare, with few reports in the literature. We describe possible pathomechanisms and propose treatment options for cases of delayed diagnosis of osteoarticular infections with Clostridium spp

Demonstrating the clinical pharmacist's activity: validation of an intervention oriented classification system

Background Clinical pharmacists are increasingly involved in detecting and solving drug-related problems. To document their performance, a convenient tool to code pharmaceutical interventions in daily practice is desirable. The Swiss Society of Public Health Administration and Hospital Pharmacists (GSASA) proposed to implement a new classification system for pharmaceutical interventions. Objectives To develop and validate a classification system for pharmaceutical interventions and to compare it with the well-established Pharmaceutical Care Network Europe (PCNE) system. Setting Rehabilitation clinic, geriatric and orthopaedic wards of a 427-bed teaching hospital. Methods Development of the GSASA classification started with expert panel discussions and the validation of the first version (GSASA V1). To assess appropriateness, interpretability, and validity, clinical pharmacists documented during a 6-week period all interventions using GSASA V1 and PCNE version 6.2 (V6.2). Acceptability and feasibility were tested by an 8-item questionnaire with 5-point Likert scale (1 = strongly disagree, 5 = strongly agree), and inter-rater reliability (Fleiss-Kappa coefficients κ) was determined. After revision, the second version (V2) was assessed again for reliability. Mean outcome measures User's agreement/satisfaction, comprehensiveness/reliability of the classification system. Results The GSASA V1 includes 4 categories and 35 subcategories. Of 115 interventions classified with GSASA V1, 93 (80.9 %) could be completely classified in all categories. This explains that 3 of 6 users could be not satisfied with the comprehensiveness of GSASA V1 (mean user agreement 2.7 ± 0.8). The questionnaire showed that all users could find GSASA V1 (4.0 ± 0.0) easier to use than PCNE V6.2 (3.0 ± 0.9). Users were generally satisfied with the GSASA V1 (3.5 ± 0.8), especially with the adequate time expenditure (4.0 ± 0.7). Inter-rater reliability and acceptability of GSASA V1 were comparable to those of the PCNE V6.2. The agreement among the GSASA V1 users was substantial for the categories 'problem' (κ = 0.66), 'intervention' (κ = 0.74), and 'outcome' (κ = 0.63), while moderate agreement for the category 'cause' was obtained (κ = 0.53). The final system GSASA V2 includes 5 categories (addition of 'type of problem') and 41 subcategories. Total inter-rater reliability was moderate (κ = 0.52). Conclusion The GSASA classification system appeared to be reliable and promising for documentation of pharmaceutical interventions in daily practice (practical and less time-consuming). The system is validated in terms of appropriateness, interpretability, validity, acceptability, feasibility, and reliability